High-throughput, real-time monitoring of engineered skeletal muscle function using magnetic sensing

Abstract

Engineered muscle tissues represent powerful tools for examining tissue level contractile properties of skeletal muscle. However, limitations in the throughput associated with standard analysis methods limit their utility for longitudinal study, high throughput drug screens, and disease modeling. Here we present a method for integrating 3D engineered skeletal muscles with a magnetic sensing system to facilitate non-invasive, longitudinal analysis of developing contraction kinetics. Using this platform, we show that engineered skeletal muscle tissues derived from both induced pluripotent stem cell and primary sources undergo improvements in contractile output over time in culture. We demonstrate how magnetic sensing of contractility can be employed for simultaneous assessment of multiple tissues subjected to different doses of known skeletal muscle inotropes as well as the stratification of healthy versus diseased functional profiles in normal and dystrophic muscle cells. Based on these data, this combined culture system and magnet-based contractility platform greatly broadens the potential for 3D engineered skeletal muscle tissues to impact the translation of novel therapies from the lab to the clinic.

Mimicking sarcolemmal damage in vitro: a contractile 3D model of skeletal muscle for drug testing in Duchenne muscular dystrophy

“…By mimicking the pathophysiological rupture of the muscle membrane through replicating repeated contraction and relaxation cycles, this model improves the current state-of-the-art dystrophic 3D cell culture tools. Recent studies have described 3D DMD models using different patient-derived cell types, including primary [52] and immortalized [53] myoblasts or induced pluripotent stem cells (iPSCs) [54,55], as well as isogenic diseased cell lines from healthy iPSCs and differentiated to myoblasts [56] or cardiomyocytes [57]. While some studies did not focus on the contractile activity in response to EPS [54], others conducted an electrophysiological analysis [58] or identified de novo dystrophin revertant fibers but were not able to detect sarcolemmal damage [53].…”

“…While some studies did not focus on the contractile activity in response to EPS [54], others conducted an electrophysiological analysis [58] or identified de novo dystrophin revertant fibers but were not able to detect sarcolemmal damage [53]. Functional DMD phenotypes have been reported in various studies, including contractile weaknesses [52], contractile and calcium transient defects [57], reduced twitch tension and prolonged contraction and relaxation times [55], as well as lower forces and contractile velocities [56]. However, the DMD 3D model developed in this work showcased the ability to not only functionally induce sarcolemmal damage but also to predict the response of potential DMD therapeutic drugs.…”

Real‐time and Multichannel Measurement of Contractility of hiPSC‐Derived 3D Skeletal Muscle using Fiber Optics‐Based Sensing

“…[19,55] To our knowledge, this sensitivity is unmatched even by more recent approaches. [56] The Cuore was developed having clearly in mind the unsettled needs emerging in the field of cardiac and skeletal muscle research. From basic biology studies to drug testing and screening, the hardware here presented allows precise and reproducible analysis of the functionality of in vitro 3D contractile tissues.…”

“…After 3 days (D3), half the volume of DM#1 was replaced by DM#2, which formulation differs from DM#1 by the concentration of KOSR increasing from 1% to 20%, which is supposed to improve contractility as suggested by previous studies. [55,56] After this point, half medium was refreshed every other day for the rest of the experiment.…”

A high-throughput 3D cantilever array to model airway smooth muscle hypercontractility in asthma

“…Elastomer-based cantilever strain gauges have emerged as a powerful means to model gross multi-cellular contractile forces in 3D at both the macro and microscales 14 . An improvement in the design of these tools is the inverted "hanging" style of construction whereby tissues are suspended from cantilevers that are mounted to the top of the cell culture multi-well plate which greatly simplifies tissue casting and improves imaging resolution 15,16 . These bioengineered tissue systems however are often lower in total throughput than highly multiplexed experiments require and utilize formats incompatible with existing microwell plate-based laboratory robotics and instrumentation 17 .…”